Nano-liquid chromatography (nanoLC) can resolve highly complex, otherwise intractable biological mixtures, and offer high sensitivity due to its enhanced resolving power. As a result when coupled to mass spectrometry, nanoLC can address complex proteomic problems such as molecular interactions, ion structures, quantitation, and kinetics. Consequently nanoLC is a necessity for biological laboratories. But unfortunately the technique is alarmingly underutilized as it suffers from limitations including reproducibility issues created by column bed inconsistencies, difficulty making nano-fluidic connections, difficulty troubleshooting system malfunctions, and the technique is labor intensive. CorSolutions is developing a robust, user-friendly, "plug- and-play" nano-LC electrospray device that will address all of the limitations of the nanoLC technique, except for the irreproducible column performance issues. These column-to-column inconsistencies arise from particle packing variations within the packed beds introduced during manufacturing, which result in performance variations and therefore limit the technology from pervasive use in the biological and clinical diagnostic fields. CorSolutions will develop automated column packing and automated, optical packed-bed inspection technology for manufacturing packed columns for nanofluidic applications. This technology will allow for both automated manufacture and subsequent inspection of the columns produced. The inspection technology will monitor light transmission through a column bed, detecting any inconsistencies in particle packing such as voids. This approach will offer consistently packed columns with reproducible column performance. Furthermore this manufacturing approach will reduce the cost of nanoLC columns, as extensive quality control testing will no longer be required. Reproducible column performance and lower column cost, together with the "plug-and-play" nanoLC system CorSolutions is developing, will dramatically reduce the complexity of nanoLC, allowing pervasive use of the technology. Also, this manufacturing approach, which could offer inexpensive, one-time use columns, may allow for nanoLC to enter the field of clinical diagnostics. The goals of the Phase I SBIR will be to first construct an automated column packing prototype, which will create columns with reproducibly packed beds and will contain an integrated photoelectric sensor to terminate the packing process when the column bed reaches a desired length. The second goal will be to construct an automated optical packed bed inspection prototype with pattern recognition software to identify bed defects. The final goal is to evaluate the prototypes by assessing column quality. This will include manufacturing 30 columns, assessing inter- and intra-column performance variables, and comparing the data to that of commercial columns. Our long term objective is to provide a novel approach for the manufacture of reproducibly performing, low cost nanofluidic columns. This will allow nanoLC to become an integral part of biological laboratories, and allow biologists to benefit from its highly informative data. PUBLIC HEALTH RELEVANCE: Nano-liquid chromatography mass spectrometry (nanoLC/MS) is a powerful technique for proteomic and glycomic studies which is able to not only identify proteins and glycans from very complex mixtures, but is also able to structurally characterize differences between the components, which is particularly important for post-translational modifications. However, nanofluidic technologies have been slow to commercialize due to lack of advanced and cost effective manufacturing approaches. Furthermore the existing manufacturing processes have not allowed for consistent column-to-column performance in conventional nanoLC, which is serving as a barrier to widespread acceptance of the technique. Therefore technology development allowing for the manufacture of columns with enhanced, reproducible performance as well as reduced column cost, will permit the powerful nanoLC technique to infiltrate all biological laboratories, which could enable scientific discoveries and greater understanding of complex biological systems.